*LB(Luria-Bertani) broth: 1% Trytone, 1% NaCl, 0.5% Yeast extract in the ddH2O *LB agar plate:LB broth add 1.5% agar.If you want to add selection marker example for ampicillin or chloramphenicol. To culture E.coli, ampicillin is added 100 mg/ml for usage, chloramphenicol is added 30 mg/ml for usage, kanamycin is added 70 mg/ml for usage
*Solution I: 50 mM glucose、25 mM Tris-HCl (pH 8.0)、10 mM EDTA *Solution II:0.2N NaOH and 1% SDS(Sodium dodecyl sulfate ) mix *Solution III: 5 M potassium acetate (pH4.8), 11.5% glacial acetic acid *RNase A(10 mg/ml) *Phenol chloroform *Chloroform *95% Ethanol *Tris-EDTA(TE)buffer:10 mM Tris-Cl(pH 7.5) 1 mM EDTA *pBluescript SK(-) *BBa_K1218011 *BBa_K914003 *BBa_I13521
*Quick screening buffer(Tris-Cl pH 12.6, 3% SDS) *Phenol chloroform *Chloroform
*T4 ligase *10X ligation buffer *EcoRI *PstI *SpeI *XbaI *BSA 0.1%
*DH5a *JM101 *0.8% and 2% agarose gel *95% Ethanol *6X loading dye *1.5 ml centrifuge tube *250 ml flask *M13KO7(1*1011 pfu/ml)from NEW ENGLAND BioLabs
*Top DNA polymerase *dNTP(dATP, dCTP, dGTP, dTTP) 250 mM *Tris-HCl(pH9.0) 10 mM *KCl 30 mM *MgCl2 1.5 mM
*VCM-620, laminal flow *Biofuge pico, Heraeus * OS1500-H, TSK Oribital Shaking Iucubator * Biowave * Vortex-GENE 2 * Autoclave AS-1060L * GenAmp PCR System 2400, Applied Biosystems *BU-420, YIHDER *5804R, eppendorf Centrifuge
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